Department of Chemical Engineering , University of Washington , Seattle , Washington 98195 , United States.
Anal Chem. 2018 Dec 18;90(24):14635-14642. doi: 10.1021/acs.analchem.8b05137. Epub 2018 Nov 28.
The performance of surface plasmon resonance (SPR)-based bacterial biosensors is often compromised as a result of diffusion-limited mass transport of bacteria to the sensing surface. In this work, dually functional interdigitated electrodes (IDEs) were developed to sustain SPR and increase bacterial mass transport through external application of dielectrophoresis (DEP). IDEs were defined into 50 nm Au films with fixed electrode gaps ( E = 5 μm) and varied electrode widths ( E = 10, 20, and 100 μm), referred to as interdigitated SPR (iSPR) chips. The iSPR chips with E = 100 μm effectively supported SPR, with comparable sensitivity to those of conventional SPR chips. The surfaces of iSPR chips ( E = 100 μm) were modified with mannose to target the FimH adhesin of Escherichia coli and increase cellular adhesion. An LOD of ∼3.0 × 10 CFU/mL E. coli was achieved on mannosylated iSPR chips under positive-DEP conditions, which is about a 5 order of magnitude improvement compared with those of mannosylated conventional SPR chips without DEP. Furthermore, secondary antibody amplification enabled selective enhancement of dilute (10 CFU/mL) E. coli suspensions, whereas no amplification was observed for concentrated (10 CFU/mL) nontarget ( Staphylococcus epidermidis) bacterial suspensions. The results presented here indicate the great potential of the incorporation of DEP into SPR biosensors for rapid, sensitive, and specific detection of bacteria with broad applications in areas of biomedical diagnostics, environmental monitoring, food safety, and homeland security.
表面等离子体共振(SPR)基细菌生物传感器的性能通常会受到细菌向传感表面扩散限制的影响。在这项工作中,开发了双重功能的叉指电极(IDE),通过施加外部介电泳(DEP)来维持 SPR 并增加细菌的质量传输。IDE 用固定的电极间隙(E=5μm)和变化的电极宽度(E=10、20 和 100μm)定义为叉指 SPR(iSPR)芯片。E=100μm 的 iSPR 芯片有效地支持 SPR,其灵敏度与传统 SPR 芯片相当。用甘露糖修饰 iSPR 芯片(E=100μm)表面以靶向大肠杆菌的 FimH 粘附素并增加细胞粘附。在正 DEP 条件下,甘露糖化的 iSPR 芯片上实现了约 3.0×10 CFU/mL 大肠杆菌的 LOD,与没有 DEP 的甘露糖化传统 SPR 芯片相比,提高了约 5 个数量级。此外,二次抗体放大使稀释(10 CFU/mL)大肠杆菌悬浮液的选择性增强,而对于浓缩(10 CFU/mL)非目标(表皮葡萄球菌)细菌悬浮液则没有观察到放大。这里呈现的结果表明,将 DEP 纳入 SPR 生物传感器中具有很大的潜力,可用于快速、灵敏和特异性地检测细菌,在生物医学诊断、环境监测、食品安全和国土安全等领域具有广泛的应用。